Skip to main content
SpringerLink
Log in

Protein-Coated Microcrystals, Combi-Protein-Coated Microcrystals, and Cross-Linked Protein-Coated Microcrystals of Enzymes for Use in Low-Water Media

  • Protocol
  • First Online:
Enzyme Stabilization and Immobilization

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1504))

Abstract

Protein-coated microcrystals (PCMC) are a high-activity preparation of enzymes for use in low-water media. The protocols for the preparation of PCMCs of Subtilisin Carlsberg and Candida antarctica lipase B (CAL B) are described. The combi-PCMC concept is useful both for cascade and non-cascade reactions. It can also be beneficial to combine two different specificities of a lipase when the substrate requires it. Combi-PCMC of CALB and Palatase used for the conversion of coffee oil present in spent coffee grounds to biodiesel is described. Cross-linked protein-coated microcrystals (CL-PCMC) in some cases can give better results than PCMC. Protocols for the CLPCMC of Subtilisin Carlsberg and Candida antarctica lipase B (CAL B) are described. A discussion of their applications is also provided.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 109.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 159.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Klibanov AM (2001) Improving enzymes by using them in organic solvents. Nature 409:241–246

    Article  CAS  PubMed  Google Scholar 

  2. Mattiasson B, Adlercreutz P (1991) Tailoring the microenvironment of enzymes in water-poor systems. Trends Biotechnol 9:394–398

    Article  CAS  PubMed  Google Scholar 

  3. Gupta MN (ed) (2000) Non-aqueous enzymology. Springer, Heidelberg

    Google Scholar 

  4. Carrea G, Riva S (2000) Properties and synthetic applications of enzymes in organic solvents. Angew Chem Int Ed 39:2226–2254

    Article  CAS  Google Scholar 

  5. Carrea G, Riva S (2008) Medium engineering. In: Gotor V, Alfonso I, Garcia-Urdiales E (eds) Asymmetric organic synthesis with enzymes. Wiley-VCH, Weinheim, pp 3–20

    Google Scholar 

  6. Zaks A, Klibanov AM (1985) Enzyme-catalyzed processes in organic solvents. Proc Natl Acad Sci U S A 82:3192–3196

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Gupta MN (1992) Enzyme function in organic solvents. Eur J Biochem 203:25–32

    Article  CAS  PubMed  Google Scholar 

  8. Dordick JS (1992) Designing enzymes for use in organic solvents. Biotechnol Prog 8:259–267

    Article  CAS  PubMed  Google Scholar 

  9. Kreiner M, Moore BD, Parkar MC (2001) Enzyme-coated micro-crystals: a 1-step method for high activity biocatalyst preparation. Chem Commun 12:1096–1097

    Article  Google Scholar 

  10. Mukherjee J, Mishra P, Gupta MN (2015) Urea treated subtilisin as a biocatalyst for transformations in organic solvents. Tetrahedron Lett 56:1976–1981

    Article  CAS  Google Scholar 

  11. Roy I, Gupta MN (2004) Preparation of highly active alpha-chymotrypsin for catalysis in organic media. Bioorg Med Chem Lett 14:2191–2193

    Article  CAS  PubMed  Google Scholar 

  12. Solanki K, Gupta MN (2008) Optimizing biocatalyst design for obtaining high transesterification activity by a-chymotrypsin in non-aqueous media. Chem Cent J 2:1–7

    Article  Google Scholar 

  13. Majumder AB, Gupta MN (2011) Increasing catalytic efficiency of Candida rugosa lipase for the synthesis of tert-alkyl butyrates in low water media. Biocatal Biotrasform 29:238–245

    Article  CAS  Google Scholar 

  14. Solanki K, Gupta MN, Halling PJ (2012) Examining structure-activity correlations of some high activity enzyme preparations for low water media. Bioresour Technol 115:147–151

    Article  CAS  PubMed  Google Scholar 

  15. Roy I, Gupta MN (2004) Hydrolysis of starch by a mixture of glucoamylase and pullulanase entrapped individually in calcium alginate beads. Enzym Microb Technol 34:26–32

    Article  CAS  Google Scholar 

  16. Rocha-Martín J, Rivas B, Muñoz R, Guisán JM, López-Gallego F (2012) Rational co-immobilization of bi-enzyme cascades on porous supports and their applications in bio-redox reactions with in situ recycling of soluble cofactors. ChemCatChem 4:1279–1288

    Article  Google Scholar 

  17. Banerjee A, Singh V, Solanki K, Mukherjee J, Gupta MN (2013) Combi-protein coated microcrystals of lipases for production of biodiesel from oil from spent coffee grounds. Sust Chem Processes 1:1–14

    Article  CAS  Google Scholar 

  18. Fernandez-Lorente G, Fernandez-Lafuente R, Armisen P, Sabuquillo P, Mateo C, Guisan JM (2000) Engineering of enzymes via immobilization and post immobilization techniques: preparation of enzyme derivatives with improved stability in organic media. In: Gupta MN (ed) Methods in non-aqueous enzymology. Birkhauser, Basel

    Google Scholar 

  19. Mateo C, Grazu V, Palomo JM, Lopez-Gallego F, Fernandez-Lafuente R, Guisan JM (2007) Immobilization of enzymes on heterofunctional epoxy supports. Nat Protoc 2:1022–1033

    Article  CAS  PubMed  Google Scholar 

  20. Shah S, Sharma A, Gupta MN (2008) Cross-linked protein-coated microcrystals as biocatalysts in non-aqueous solvents. Biocatal Biotransform 26:266–271

    Article  CAS  Google Scholar 

  21. Raita M, Laothanachareon T, Champreda V, Laosiripojana N (2011) Biocatalytic esterification of palm oil fatty acids for biodiesel production using glycine-based cross-linked protein coated microcrystalline lipase. J Mol Catal B Enzym 73:74–79

    Article  CAS  Google Scholar 

  22. Gupta MN, Shah S, Sharma A (2006) A method for preparation of cross-linked protein coated microcrystals (Appln. No. 2046/DEL/2006 dated 18-9-2006)

    Google Scholar 

  23. Kapoor M, Gupta MN (2012) Obtaining monoglycerides by esterification of glycerol with palmitic acid using some high activity preparations of Candida antarctica lipase B. Process Biochem 47:503–508

    Article  CAS  Google Scholar 

  24. Halling PJ (1992) Salt hydrates for water activity control with biocatalysts in organic media. Biotechnol Tech 6:271–276

    Article  CAS  Google Scholar 

  25. Schoevaart R, Wolbers MW, Golubovic M, Ottens M, Kieboom AP, van Rantwijk F, van der Wielen LA, Sheldon RA (2004) Preparation, optimization, and structures of cross-linked enzyme aggregates (CLEAs). Biotechnol Bioeng 87:754–762

    Article  CAS  PubMed  Google Scholar 

  26. Kumari V, Shah S, Gupta MN (2007) Preparation of biodiesel by lipase-catalyzed transesterification of high free fatty acid containing oil from madhuca indica. Energ Fuels 21:368–372

    Article  CAS  Google Scholar 

  27. Chiaradia V, Paroul N, Cansian RL, Júnior CV, Detofol MR, Lerin LA, Oliveira JV, Oliveira D (2012) Synthesis of eugenol esters by lipase-catalyzed reaction in solvent-free system. Appl Biochem Biotechnol 168:742–751

    Article  CAS  PubMed  Google Scholar 

  28. Solanki K, Gupta MN (2011) A chemically modified lipase preparation for catalyzing the transesterification reaction in even highly polar organic solvents. Bioorg Med Chem Lett 21:2934–2936

    Article  CAS  PubMed  Google Scholar 

  29. Kreiner M, Parker MC (2004) High-activity biocatalysts in organic media: solid-state buffers as the immobilisation matrix for protein-coated microcrystals. Biotechnol Bioeng 87:24–33

    Article  CAS  PubMed  Google Scholar 

  30. Zheng J, Xu L, Liu Y, Zhang X, Yan Y (2012) Lipase-coated K2SO4 micro-crystals: preparation, characterization, and application in biodiesel production using various oil feedstocks. Bioresour Technol 110:224–231

    Article  CAS  PubMed  Google Scholar 

  31. Wu JC, Yang JX, Zhang SH, Chow Y, Talukder MR, Choi WJ (2009) Activity, stability and enantioselectivity of lipase-coated microcrystals of inorganic salts in organic solvents. Biocatal Biotransform 27:283–289

    Article  CAS  Google Scholar 

  32. Murdan S, Somavarapu S, Ross AC, Alpar HO, Parker MC (2005) Immobilisation of vaccines onto micro-crystals for enhanced thermal stability. Int J Pharm 296:117–121

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by funds obtained from Department of Science and Technology [Grant No.: SR/SO/BB-68/2010] and Department of Biotechnology [Grant No.: BT/PR14103/BRB/10/808/2010], both Government of India organizations. Finally, we thank past members of our research group; Dr. Aparna Sharma, Dr. Shweta Shah, Dr. Kusum Solanki, Dr. Manali Kapoor, Veena Singh, and Aditi Banerjee, whose data have been described/quoted in this chapter. We would also like to acknowledge the collaboration in this area with Prof. Peter J. Halling (University of Strathclyde, Glasgow, UK) funded by UKIERI, UK (2008–2012).

Author information

Authors and Affiliations

  1. Chemistry Department, Indian Institute of Technology Delhi, Hauz Khas, New Delhi, 110 016, India

    Joyeeta Mukherjee

  2. Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology Delhi, Hauz Khas, New Delhi, 110 016, India

    Munishwar N. Gupta

Authors
  1. Joyeeta Mukherjee
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Munishwar N. Gupta
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Munishwar N. Gupta .

Editor information

Editors and Affiliations

  1. Department of Chemistry, and Department of Materials Science and Engineering, University of Utah, Salt Lake City, Utah, USA

    Shelley D. Minteer

Rights and permissions

Reprints and permissions

Copyright information

© 2017 Springer Science+Business Media New York

About this protocol

Cite this protocol

Mukherjee, J., Gupta, M.N. (2017). Protein-Coated Microcrystals, Combi-Protein-Coated Microcrystals, and Cross-Linked Protein-Coated Microcrystals of Enzymes for Use in Low-Water Media. In: Minteer, S. (eds) Enzyme Stabilization and Immobilization. Methods in Molecular Biology, vol 1504. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6499-4_10

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-6499-4_10

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6497-0

  • Online ISBN: 978-1-4939-6499-4

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation